骨髓间充质干细胞条件培养基加入罗格列酮后促进内皮祖细胞的增殖与迁移

Proliferation and migration of endothelial progenitor cells promoted by bone marrow mesenchymal stem cells conditioned medium with rosiglitazone

背景:研究表明间充质干细胞可参与修复糖尿病引发的创面损伤,但高糖环境明显抑制了间充质干细胞的功能与移植效果。目的:观察罗格列酮干预的骨髓间充质干细胞条件培养基对高糖环境下内皮祖细胞增殖、迁移的作用。方法:①取对数生长期的骨髓间充质干细胞,分3组培养,正常组加入含体积分数为10%胎牛血清的α-MEM培养基培养;高糖组加入含体积分数为10%胎牛血清、25 mmol/L葡萄糖的α-MEM培养基培养;罗格列酮组加入含体积分数为10%胎牛血清、25 mmol/L葡萄糖、10μmol/L罗格列酮的α-MEM培养基培养,培养48 h后提取培养上清作为条件培养基,ELISA法检测条件培养基中血管内皮生长因子与基质细胞衍生因子1水平。②取对数生长期的内皮祖细胞,分3组干预,对照组加入含体积分数为10%胎牛血清的EGM-2 MV培养基;模型组加入含体积分数为10%胎牛血清、30 mmol/L葡萄糖、高糖组条件培养基的EGM-2 MV培养基;实验组加入含体积分数为10%胎牛血清、30 mmol/L葡萄糖、罗格列酮组条件培养基的EGM-2 MV培养基,培养24 h后进行细胞增殖与迁移能力检测。结果与结论:①高糖组条件培养基中血管内皮生长因子与基质细胞衍生因子1水平明显低于正常组(P <0.05),罗格列酮组条件培养基中血管内皮生长因子与基质细胞衍生因子1水平均明显高于高糖组(P <0.05);②模型组内皮祖细胞的增殖与迁移能力明显低于对照组(P <0.05);实验组内皮祖细胞的增殖与迁移能力高于模型组(P <0.05);③结果提示罗格列酮干预的骨髓间充质干细胞条件培养基可促进内皮祖细胞的增殖与迁移。

BACKGROUND:Studies have shown that mesenchymal stem cells can participate in the repair of wound injury caused by diabetes,but the high glucose environment obviously inhibits the function of mesenchymal stem cells and the effect of transplantation.OBJECTIVE:To observe the effect of conditioned medium of bone marrow mesenchymal stem cells intervened by rosiglitazone on the proliferation and migration of endothelial progenitor cells in high glucose environment.METHODS:(1)The bone marrow mesenchymal stem cells from the logarithmic growth period were cultured in three groups.The normal group was cultured with alpha-MEM medium containing 10%fetal bovine serum.The high glucose group was cultured with alpha-MEM medium containing 10%fetal bovine serum and 25 mmol/L glucose.The rosiglitazone group was cultured with alpha-MEM medium containing 10%fetal bovine serum,25 mmol/L glucose and 10μmol/L rosiglitazone.After 48 hours of culture,the culture supernatant was extracted as conditioned medium.The levels of vascular endothelial growth factor and matrix cell derived factor 1 in conditioned medium were detected by ELISA.(2)The endothelial progenitor cells from the logarithmic growth period were divided into three groups.The control group was cultured with the EGM-2 MV medium containing 10%fetal bovine serum.The model group was cultured with the EGM-2 MV medium containing 10%fetal bovine serum,30 mmol/L glucose and conditioned medium of the high glucose group.The experimental group was cultured with EGM-2 MV medium containing 10%fetal bovine serum,30 mmol/L glucose and conditioned medium of the rosiglitazone group.After 24 hours of culture,the ability of cell proliferation and migration was detected.RESULTS AND CONCLUSION:(1)The levels of vascular endothelial growth factor and matrix cell derived factor 1 in the conditioned medium of high glucose group were significantly lower than that of the normal group(P<0.05).The levels of vascular endothelial growth factor and matrix cell derived factor 1 in the conditioned medium of the rosiglitazone group were significantly higher than in the high glucose group(P<0.05).(2)The proliferation and migration ability of endothelial progenitor cells in the model group was lower than that in the control group(P<0.05).The proliferation and migration ability of endothelial progenitor cells in the experimental group was higher than that in the model group(P<0.05).(3)It is suggested that the conditioned medium of rosiglitazone intervened bone marrow mesenchymal stem cells can promote the proliferation and migration of endothelial progenitor cells.